HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Weisgraber, K. H. Articles by Mahley, R. W. Search for Related Content PubMed PubMed Citation Articles by Weisgraber, K. H. Articles by Mahley, R. W. Social Bookmarking                      What's this?

Journal of Lipid Research, Vol. 21, 316-325, March 1980
Copyright © 1980 by Lipid Research, Inc.

Subfractionation of human high density lipoproteins by heparin-Sepharose affinity chromatography

Karl H. Weisgraber and Robert W. Mahley

Gladstone Foundation Laboratories for Cardiovascular Disease, University of California, San Francisco, CA 94140 and National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20205

A reproducible and quantitative subfractionation of human high density lipoproteins (HDL) by heparin-Sepharose affinity chromatography has been developed. Two elution methods (A and B) were used to subfractionate HDL₂ (d 1.063-1.125 g/ml) or total HDL (d 1.063-1.21 g/ml). Method A separated HDL₂ into three subclasses, each with distinct chemical properties and in vitro metabolic characteristics. The first subclass, referred to as HDL₂-without E, passed through the affinity column unretarded and represented approximately 85% of the HDL₂ lipoprotein protein. HDL₂-without E contained the A-I, A-II, and C apoproteins which characterize typical HDL. The second subclass eluted from the column (7-10% of the protein) contained, in addition to the A-I and A-II apoproteins, the E and (E—A-II) apoproteins, and was designated as HDL₂-with E. The B apoprotein was the major protein component of the third subclass eluted from the column (ßbeta; lipoproteins). The ßbeta; subclass accounted for approximately 3-8% of the HDL₂ protein and was similar to Lp(a) in composition and size. Method B further subdivided the ßbeta; subclass into two fractions (ßbeta;₁ and ßbeta;₂) with slightly different electrophoretic mobilities. The various heparin-Sepharose subclasses were further characterized by their ability to compete with ¹²⁵I-labeled low density lipoproteins (LDL) for binding to cell surface receptor sites of fibroblasts. By virtue of the presence of the E apoprotein, HDL₂-with E competed effectively with ¹²⁵I-labeled LDL for binding to the cell surface receptors, whereas HDL₂-without E were ineffective in competing with LDL. The ßbeta; subclass possessed binding capability similar to that of LDL. Subfractionation of HDL by heparin-Sepharose affinity column chromatography provides an attractive alternative to methods based solely on ultracentrifugation, in that it subfractionates HDL into subclasses with differing apoprotein contents that impart distinct metabolic characteristics to each class.—Weisgraber, K. H., and R. W. Mahley. Subfractionation of human high density lipoproteins by heparin-Sepharose affinity chromatography.

Supplementary key words apoprotein E • HDL subclasses • apo(E—A-II) complex • Lp(a) • lipoprotein cell receptors

Submitted on August 29, 1979
Revised on November 20, 1979

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				C. Xie, S. D. Turley, and J. M. Dietschy ABCA1 plays no role in the centripetal movement of cholesterol from peripheral tissues to the liver and intestine in the mouse J. Lipid Res., July 1, 2009; 50(7): 1316 - 1329. [Abstract] [Full Text] [PDF]

				B. Liu, C. Xie, J. A. Richardson, S. D. Turley, and J. M. Dietschy Receptor-mediated and bulk-phase endocytosis cause macrophage and cholesterol accumulation in Niemann-Pick C disease J. Lipid Res., August 1, 2007; 48(8): 1710 - 1723. [Abstract] [Full Text] [PDF]

				P. A. C. McPherson, I. S. Young, B. McKibben, and J. McEneny High density lipoprotein subfractions: isolation, composition, and their duplicitous role in oxidation J. Lipid Res., January 1, 2007; 48(1): 86 - 95. [Abstract] [Full Text] [PDF]

				M. C. Cheung and J. J. Albers Active plasma phospholipid transfer protein is associated with apoA-I- but not apoE-containing lipoproteins J. Lipid Res., June 1, 2006; 47(6): 1315 - 1321. [Abstract] [Full Text] [PDF]

				C. Xie, J. A. Richardson, S. D. Turley, and J. M. Dietschy Cholesterol substrate pools and steroid hormone levels are normal in the face of mutational inactivation of NPC1 protein J. Lipid Res., May 1, 2006; 47(5): 953 - 963. [Abstract] [Full Text] [PDF]

				S. Siggins, M. Jauhiainen, V. M. Olkkonen, J. Tenhunen, and C. Ehnholm PLTP secreted by HepG2 cells resembles the high-activity PLTP form in human plasma J. Lipid Res., September 1, 2003; 44(9): 1698 - 1704. [Abstract] [Full Text] [PDF]

				D. L. Macdonald, T. L. Terry, L. B. Agellon, P. N. Nation, and G. A. Francis Administration of Tyrosyl Radical-Oxidized HDL Inhibits the Development of Atherosclerosis in Apolipoprotein E-Deficient Mice Arterioscler Thromb Vasc Biol, September 1, 2003; 23(9): 1583 - 1588. [Abstract] [Full Text] [PDF]

				H. Y. Choi, B. Karten, T. Chan, J. E. Vance, W. L. Greer, R. A. Heidenreich, W. S. Garver, and G. A. Francis Impaired ABCA1-dependent Lipid Efflux and Hypoalphalipoproteinemia in Human Niemann-Pick type C Disease J. Biol. Chem., August 29, 2003; 278(35): 32569 - 32577. [Abstract] [Full Text] [PDF]

				B. Karten, D. E. Vance, R. B. Campenot, and J. E. Vance Trafficking of Cholesterol from Cell Bodies to Distal Axons in Niemann Pick C1-deficient Neurons J. Biol. Chem., January 31, 2003; 278(6): 4168 - 4175. [Abstract] [Full Text] [PDF]

				C. Xie, L. A. Woollett, S. D. Turley, and J. M. Dietschy Fatty acids differentially regulate hepatic cholesteryl ester formation and incorporation into lipoproteins in the liver of the mouse J. Lipid Res., September 1, 2002; 43(9): 1508 - 1519. [Abstract] [Full Text] [PDF]

				F. Rinninger, M. Brundert, I. Brosch, N. Donarski, R. M. Budzinski, and H. Greten Lipoprotein lipase mediates an increase in selective uptake of HDL-associated cholesteryl esters by cells in culture independent of scavenger receptor BI J. Lipid Res., November 1, 2001; 42(11): 1740 - 1751. [Abstract] [Full Text] [PDF]

				S. Koch, N. Donarski, K. Goetze, M. Kreckel, H.-J. Stuerenburg, C. Buhmann, and U. Beisiegel Characterization of four lipoprotein classes in human cerebrospinal fluid J. Lipid Res., July 1, 2001; 42(7): 1143 - 1151. [Abstract] [Full Text] [PDF]

				C. Xie, S. D. Turley, and J. M. Dietschy Centripetal cholesterol flow from the extrahepatic organs through the liver is normal in mice with mutated Niemann-Pick type C protein (NPC1) J. Lipid Res., August 1, 2000; 41(8): 1278 - 1289. [Abstract] [Full Text]

				C. Xie, S. D. Turley, and J. M. Dietschy Cholesterol accumulation in tissues of the Niemann-Pick type C mouse is determined by the rate of lipoprotein-cholesterol uptake through the coated-pit pathway in each organ PNAS, October 12, 1999; 96(21): 11992 - 11997. [Abstract] [Full Text] [PDF]

				K. Fluiter, W. Sattler, M. C. De Beer, P. M. Connell, D. R. van der Westhuyzen, and T. J. C. van Berkel Scavenger Receptor BI Mediates the Selective Uptake of Oxidized Cholesterol Esters by Rat Liver J. Biol. Chem., March 26, 1999; 274(13): 8893 - 8899. [Abstract] [Full Text] [PDF]

				D. V.-v. Bruggen, I. Kalkman, T. van Gent, A. van Tol, and H. Jansen Induction of Adrenal Scavenger Receptor BI and Increased High Density Lipoprotein-Cholesteryl Ether Uptake by in Vivo Inhibition of Hepatic Lipase J. Biol. Chem., November 27, 1998; 273(48): 32038 - 32041. [Abstract] [Full Text] [PDF]

				W.-Q. Wang, D. L. Merriam, A. S. Moses, and G. A. Francis Enhanced Cholesterol Efflux by Tyrosyl Radical-oxidized High Density Lipoprotein Is Mediated by Apolipoprotein AI-AII Heterodimers J. Biol. Chem., July 10, 1998; 273(28): 17391 - 17398. [Abstract] [Full Text] [PDF]

				F. Rinninger, T. Kaiser, W. A. Mann, N. Meyer, H. Greten, and U. Beisiegel Lipoprotein lipase mediates an increase in the selective uptake of high density lipoprotein-associated cholesteryl esters by hepatic cells in culture J. Lipid Res., July 1, 1998; 39(7): 1335 - 1348. [Abstract] [Full Text]

				K. Fluiter, D. R. van der Westhuijzen, and T. J. C. van Berkel In Vivo Regulation of Scavenger Receptor BI and the Selective Uptake of High Density Lipoprotein Cholesteryl Esters in Rat Liver Parenchymal and Kupffer Cells J. Biol. Chem., April 3, 1998; 273(14): 8434 - 8438. [Abstract] [Full Text] [PDF]

				E. I. P. de Chaves, A. E. Rusinol, D. E. Vance, R. B. Campenot, and J. E. Vance Role of Lipoproteins in the Delivery of Lipids to Axons during Axonal Regeneration J. Biol. Chem., December 5, 1997; 272(49): 30766 - 30773. [Abstract] [Full Text] [PDF]

				P. N. Duchateau, C. R. Pullinger, R. E. Orellana, S. T. Kunitake, J. Naya-Vigne, P. M. O'Connor, M. J. Malloy, and J. P. Kane Apolipoprotein L, a New Human High Density Lipoprotein Apolipoprotein Expressed by the Pancreas. IDENTIFICATION, CLONING, CHARACTERIZATION, AND PLASMA DISTRIBUTION OF APOLIPOPROTEIN L J. Biol. Chem., October 10, 1997; 272(41): 25576 - 25582. [Abstract] [Full Text] [PDF]

				P. C.N. Rensen and T. J.C. van Berkel Apolipoprotein E Effectively Inhibits Lipoprotein Lipase-mediated Lipolysis of Chylomicron-like Triglyceride-rich Lipid Emulsions in Vitro and in Vivo J. Biol. Chem., June 21, 1996; 271(25): 14791 - 14799. [Abstract] [Full Text] [PDF]

				H.-O. Mowri, J. R. Patsch, A. M. Gotto Jr, and W. Patsch Apolipoprotein A-II Influences the Substrate Properties of Human HDL2 and HDL3 for Hepatic Lipase Arterioscler Thromb Vasc Biol, June 1, 1996; 16(6): 755 - 762. [Abstract] [Full Text]

				T. O'Brien, T. T. Nguyen, B. J. Hallaway, D. Hodge, K. Bailey, D. Holmes, and B. A. Kottke The Role of Lipoprotein A-I and Lipoprotein A-I/A-II in Predicting Coronary Artery Disease Arterioscler Thromb Vasc Biol, February 1, 1995; 15(2): 228 - 231. [Abstract] [Full Text]

				R. Mahley Apolipoprotein E: cholesterol transport protein with expanding role in cell biology Science, April 29, 1988; 240(4852): 622 - 630. [Abstract] [PDF]

				M. Ignatius, E. Shooter, R. Pitas, and R. Mahley Lipoprotein uptake by neuronal growth cones in vitro Science, May 22, 1987; 236(4804): 959 - 962. [Abstract] [PDF]